title: Reactive oxygen species play a critical role in collagen-induced platelet activation via SHP-2 oxidation.
abstract: OBJECTIVE
  The collagen-stimulated generation of reactive oxygen species (ROS) regulates signal transduction in platelets, although the mechanism is unclear. The major targets of ROS include protein tyrosine phosphatases (PTPs). ROS-mediated oxidation of the active cysteine site in PTPs abrogates the PTP catalytic activity. The aim of this study was to elucidate whether collagen-induced ROS generation leads to PTP oxidation, which promotes platelet stimulation.
  
  
  RESULTS
  SH2 domain-containing PTP-2 (SHP-2) is oxidized in platelets by ROS produced upon collagen stimulation. The oxidative inactivation of SHP-2 leads to the enhanced tyrosine phosphorylation of spleen tyrosine kinase (Syk), Vav1, and Bruton's tyrosine kinase (Btk) in the linker for the activation of T cells signaling complex, which promotes the tyrosine phosphorylation-mediated activation of phospholipase Cγ2 (PLCγ2). Moreover, we found that, relative to wild-type platelets, platelets derived from glutathione peroxidase 1 (GPx1)/catalase double-deficient mice showed enhanced cellular ROS levels, oxidative inactivation of SHP-2, and tyrosine phosphorylation of Syk, Vav1, Btk, and PLCγ2 in response to collagen, which subsequently led to increased intracellular calcium levels, degranulation, and integrin αIIbβ3 activation. Consistent with these findings, GPx1/catalase double-deficiency accelerated the thrombotic response in FeCl3-injured carotid arteries.
  
  
  METHODS
  The present study is the first to demonstrate that SHP-2 is targeted by ROS produced in collagen-stimulated platelets and suggests that a novel mechanism for the regulation of platelet activation by ROS is due to oxidative inactivation of SHP-2.
  
  
  CONCLUSIONS
  We conclude that collagen-induced ROS production leads to SHP-2 oxidation, which promotes platelet activation by upregulating tyrosine phosphorylation-based signal transduction.
authors.affiliation: 1 Graduate School of Pharmaceutical Sciences and College of Pharmacy, Ewha Womans University , Seoul, South Korea .
authors.forename: Ji Yong
authors.initials: JY
authors.lastname: Jang
authors.forename: Ji Hyun
authors.initials: JH
authors.lastname: Min
authors.forename: Yun Hee
authors.initials: YH
authors.lastname: Chae
authors.forename: Jin Young
authors.initials: JY
authors.lastname: Baek
authors.forename: Su Bin
authors.initials: SB
authors.lastname: Wang
authors.forename: Su Jin
authors.initials: SJ
authors.lastname: Park
authors.forename: Goo Taeg
authors.initials: GT
authors.lastname: Oh
authors.forename: Sang-Hak
authors.initials: SH
authors.lastname: Lee
authors.forename: Ye-Shih
authors.initials: YS
authors.lastname: Ho
authors.forename: Tong-Shin
authors.initials: TS
authors.lastname: Chang
mesh_terms: D000818:Animals; D003094:Collagen / Q000494:pharmacology*; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D010084:Oxidation-Reduction; D015539:Platelet Activation / Q000187:drug effects*; D054592:Protein Tyrosine Phosphatase, Non-Receptor Type 11 / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism*
chemical_list: D017382:Reactive Oxygen Species; D003094:Collagen; C516724:PTPN11 protein, human; D054592:Protein Tyrosine Phosphatase, Non-Receptor Type 11